Orchids of Azerbaijani Cemeteries

In order to explore their orchid flora, we performed surveys of 96 Azerbaijani burial places in 2018 and 2019. Altogether, 28 orchid taxa were found in 37 visited cemeteries. In the orchid diversity a remarkable pattern was observed: geographic latitude was significantly and positively related to the number of taxa and number of individuals. The most widespread and abundant orchids in Azerbaijani graveyards were Anacamptis pyramidalis and A. papilionacea (found in 23 and 8 cemeteries, respectively). Azerbaijani cemeteries can be important refuges for rare and threatened orchids, e.g., Himantoglossum formosum (three cemeteries), Ophrys sphegodes subsp. mammosa (eight), Orchis adenocheila (two), O. punctulata (three), O. stevenii (one) and Steveniella satyrioides (one). Epipactis turcica, detected in a single locality, was previously unknown to the flora of Azerbaijan. Additionally, we documented orchid tuber (salep) collection in two cemeteries.

Phytochemical Analysis, Network Pharmacology and in Silico Investigations on Anacamptis pyramidalis Tuber Extracts

Anacamptis pyramidalis (L.) Rich. forms part of the Orchidaceae family that is highly valued for its horticultural as well as therapeutic benefits. The present study set out to investigate the inhibitory activity of A. pyramidalis tubers against key biological targets for the management of type 2 diabetes, Alzheimer disease, and skin hyperpigmentation. In addition, the antioxidant potential of the extracts was also assessed using multiple methods. The detailed phytochemical profiles of the extracts were determined using high-performance liquid chromatography. Based on qualitative phytochemical fingerprint, a network pharmacology analysis was conducted as well. Parishin was identified from the water extract only, whereas gastrodin and caffeic acid derivatives were present in the methanol extract. The methanol extract exhibited high inhibitory activity against tyrosinase (69.69 mg kojic acid equivalent/g extract), α-amylase (15.76 mg acarbose equivalent/g extract), and α-glucosidase (20.07 mg acarbose equivalent/g extract). Similarly, the methanol extract showed highest antioxidant potential (22.12, 44.23, 45.56, and 29.38 mg Trolox equivalent/g extract, for 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), CUPric Reducing Antioxidant Capacity (CUPRAC), and Ferric Reducing Antioxidant Power (FRAP) assays, respectively). Finally, the results of network pharmacology analysis, besides corroborating traditional uses of plant extracts in the management of cold and flu, confirmed a direct involvement of identified phytochemicals in the observed enzyme inhibitory effects, especially against tyrosinase, α-amylase, and α-glucosidase. Furthermore, based on the results of both colorimetric assays and network pharmacology analysis related to the activity of A. pyramidalis extracts and identified phytocompounds on enzymes involved in type 2 diabetes, a docking study was conducted in order to investigate the putative interactions of oxo-dihydroxy octadecenoic acid trihydroxy octadecenoic acid against aldose reductase, peroxisome proliferator-activated receptor (PPAR)-α, dipeptidyl peptidase (DPP)-IV, and α-glucosidase. Docking analysis suggested the inhibitory activity of these compounds against the aforementioned enzymes, with a better inhibitory profile shown by oxo-dihydroxy octadecenoic acid. Overall, the present findings supported the rationale for the use of A. pyramidalis as source of bioactive metabolites and highlight, today more than ever, for the strong necessity of linkage strategy between wild resource valorization and conservation policy.

Characterization and Antioxidant Activity of Essential Oil of Four Sympatric Orchid Species

The volatile fractions from fresh inflorescences of naturally growing orchids Anacamptis coriophora (L.) R. M. Bateman, Pridgeon & M. W. Chase subsp. fragrans (Pollini), Anacamptis pyramidalis (L.) R. Ophrys holosericea (Burm.) Greuter and Serapias vomeracea (Burm. f.) B. were isolated by steam distillation and analyzed by GC/FID and GC/MS. Saturated hydrocarbons were quantified as the major constituents of the volatile fraction (47.87–81.57% of the total essential oil), of which long-chain monounsaturated hydrocarbons accounted from 9.20% to 32.04% of the total essential oil. Double bond position in linear alkenes was highlighted by dimethyl disulfide derivatization and MS fragmentation. Aldehydes (from 3.45 to 18.18% of the total essential oil), alcohols (from 0.19% to 13.48%), terpenes (from 0.98 to 2.50%) and acids (0.30 to 2.57%) were also detected. These volatiles compounds may represent a particular feature of these plant species, playing a critical role in the interaction with pollinators. DPPH assay evaluating the antioxidant activity of the essential oils was carried out, showing a dose-dependent antioxidant activity.

INVESTIGATING THE EFFECT OF SUBSTRATE, MYCORRHIZAL APPLICATION AND BULB SEPARATION ON THE GROWTH OF THE WILD ORCHID ANACAMPTIS PYRAMIDALIS

Pyramidal orchid (Anacamptis pyramidalis) is a wild terrestrial orchid widelyfound in Lebanon and the Mediterranean zone. Random collection and trade of theorchid for medicinal and edible use (salep) subjected it to a risk of extinction.Consequently, the current work aimed to propagate this orchid in vivo. Orchidbulbs were transplanted from the wild into four different soil substrates (Pinebark,Pinebark-Peat (1/1), Peat-Sand (1/1) and control: soil from the collection site)under controlled conditions of temperature and humidity. The effect of mycorrhizalapplication (MY: Yes) was tested in the different substrates except in control andorchid bulbs (son bulb and mother bulb) were planted combined (SB+MB) or afterseparation of the mother bulb (SB). Same treatments were repeated over twoconsecutive years. Results showed an earlier emergence of son bulbs (SB) grownin control substrate. Moreover, there was a significant difference in plant growthwith superiority for (SB) compared to (SB+MB) regarding plant length andelongation of first leaf except bulb dimensions (length and width) that were higherin (SB+MB). Mycorrhizal application enhanced the overall growth of plants and itseffect was the most obvious in the substrate Peat-Sand (1/1). In general, the bestgrowth of the orchid was observed at the level (SB)/(MY:Yes)/Peat-Sand (1/1).Results of the second experimental year confirmed those of the first year with animprovement of the rate of emergence by 13%. The in vivo propagation methodwas beneficial on improving the growth of A. pyramidalis ex situ and it could beadopted as an initiative for wild orchid conservation.

Shift in flowering time allows diploid and autotetraploid Anacamptis pyramidalis (Orchidaceae) to coexist by reducing competition for pollinators

How autopolyploids establish when competing with diploid progenitors is still poorly understood. Here, we investigate the effects of morphological and phenological divergence on the reproductive success of co-occurring diploid and autotetraploid individuals of Anacamptis pyramidalis to disentangle the mechanisms allowing their c-oexistence. We estimated reproductive success in terms of absolute and relative fruit production and recorded morphological and phenological traits of the two cytotypes. We also estimated correlations between traits within cytotypes and the related phenotypic selection patterns. The two cytotypes experience similar levels of reproductive success, but they differ significantly in their morphological and phenological traits. Correlations among floral traits were weaker in autotetraploids than in diploids, but among-individuals variation was higher in diploids. For most flower traits, the strength of phenotypic selection was different in the two cytotypes. We found selection on flower number in both cytotypes, whereas selection for earlier flowering time was only found in late-flowering diploids. Our results suggest that a shift in flowering time may have allowed diploid and autotetraploid A. pyramidalis individuals to reduce competition for naïve pollinators. Consequently, the two cytotypes can coexist by attaining similar levels of reproductive success and, at same time, reducing the likelihood of inter-cytotype mating.